Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms

Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms - Download this document for free, or read online. Document in PDF available to download.

Microbial Cell Factories

, 9:91

First Online: 23 November 2010Received: 30 August 2010Accepted: 23 November 2010

Abstract

BackgroundFumarase catalyzes the reversible hydration of fumarate to L-malate and is a key enzyme in the tricarboxylic acid TCA cycle and in amino acid metabolism. Fumarase is also used for the industrial production of L-malate from the substrate fumarate. Thermostable and high-activity fumarases from organisms that inhabit extreme environments may have great potential in industry, biotechnology, and basic research. The marine environment is highly complex and considered one of the main reservoirs of microbial diversity on the planet. However, most of the microorganisms are inaccessible in nature and are not easily cultivated in the laboratory. Metagenomic approaches provide a powerful tool to isolate and identify enzymes with novel biocatalytic activities for various biotechnological applications.

ConclusionsWe isolated a novel fumarase gene, fumF, from a sequence-based screen of a plasmid metagenomic library from uncultivated marine microorganisms. The properties of FumF protein may be ideal for the industrial production of L-malate under higher temperature conditions. The identification of FumF underscores the potential of marine metagenome screening for novel biomolecules.

List of abbreviations usedFumAfumarase A

FumBfumarase B

FumCfumarase C

SDS-PAGEsodium dodecyl sulfate-polyacrylamide gel electrophoresis

E. coliEscherichia coli

Ni-NTAnickel-nitrilotriacetic acid

HPLChigh performance liquid chromatography

EDTAethylenediaminetetraacetic acid

SDSsodium dodecyl sulfate

ORFopen reading frame.

Electronic supplementary materialThe online version of this article doi:10.1186-1475-2859-9-91 contains supplementary material, which is available to authorized users.

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